糖尿病神经性骨关节病的病理表现与分子发病机制的研究  

Pathological characters and molecular pathogenesis of diabetic neuropathic osteoarthropathy

作　　者：刘培珑 刁佳宇 梁景棋 张言[1] 温晓东 文嫣 郝婧灿 梁晓军[1] 张峰[3] 郝定均[5] 赵宏谋[1] Liu Peilong;Diao Jiayu;Liang Jingqi;Zhang yan;Wen Xiaodong;Wen Yan;Hao Jingcan;Liang Xiaojun;Zhang Feng;Hao Dingjun;Zhao Hongmou(Department of Foot and Ankle Surgery,Honghui Hospital of Xi'an Jiaotong University,Xi'an 710054,China;The Second Department of Cardiovascular,Shanxi Provincial People's Hospital,Xi'an 710068,China;School of Public Health,Xi'an Jiaotong University,Xi'an 710049,China;Cancer Center,the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061,China;Spine Hospital,Honghui Hospital of Xi'an Jiaotong University,Xi'an 710054,China)

机构地区：[1]西安交通大学附属红会医院足踝外科,西安710054 [2]陕西省人民医院心血管内二科,西安710068 [3]西安交通大学公共卫生学院,西安710049 [4]西安交通大学第一附属医院癌症中心,西安710061 [5]西安交通大学附属红会医院脊柱病医院,西安710054

出　　处：《中华解剖与临床杂志》2020年第1期67-73,共7页Chinese Journal of Anatomy and Clinics

基　　金：中国博士后科学基金(2017M613178);陕西省博士后科研基金(2017BSHQYXMZZ13);陕西省自然科研基金(2018JM7112)。

摘　　要：目的观察糖尿病神经性骨关节病(DNOAP)患者软骨的病理表现并探讨其发病机制。方法选取2017年3月—2018年6月西安市红会医院收治的8例DNOAP患者的胫距关节、距下关节、距舟关节的关节软骨为DNOAP组,其中男3例、女5例,年龄20~66(55.7±3.8)岁;2017年4月—2018年7月西安市红会医院因车祸或严重外伤截肢的8例患者的胫距关节、距下关节、距舟关节的关节软骨为正常对照组,其中男4例,女4例,年龄19~65(57.6±3.7)岁。取DNOAP组和正常对照组软骨样本,使用Masson染色和番红O/固绿染色观察软骨组织病理学特征,并通过电子显微镜观察软骨细胞的超微结构变化。取DNOAP组和正常对照组软骨样本进行软骨细胞培养,采用DCFH-DA探针方法通过Image-Pro Plus软件分析荧光强度检测软骨细胞内活性氧的水平,采用Western blot检测软骨细胞中核因子κB受体活化因子配体(RANKL)、白细胞介素(IL)-1β、骨保护素(OPG)、IL-6、肿瘤坏死因子(TNF)-α、聚集蛋白聚糖(aggrecan)蛋白的表达水平,采用流式细胞术检测软骨细胞凋亡率。结果DNOAP组软骨病理检查见透明软骨呈条索状排列,软骨细胞减少,软骨下骨增生,结构紊乱,软骨下骨区域大量破骨细胞形成;透射电子显微镜下见DNOAP软骨细胞的线粒体肿胀,膜结构不完整,排列紊乱,内质网严重肿胀,细胞核变大,染色质出现部分断裂,浓聚在核膜边缘。正常对照组软骨病理检查:正常软骨组织可见软骨细胞位于软骨陷窝内,软骨基质染色均匀,软骨下骨排列整齐;透射电子显微镜下:软骨细胞可见丰富的粗面内质网,细胞核内可见正常浓缩的染色质。DNOAP组软骨细胞的活性氧荧光强度为28.1±2.3,高于正常对照组的11.9±0.7,差异有统计学意义(t=19.059,P<0.01)。Western blot检测DNOAP组RANKL、TNF-α、IL-1β及IL-6蛋白的相对表达量均高于正常对照组,而OPG、aggrecan蛋白的相对表达量均低于正常�Objective To investigate the pathological characters and relative molecular pathogenesis of diabetic neuropathic osteoarthropathy(DNOAP).Methods The articular cartilages of tibial talus,subtalar joint and talo-scaphoid joint of 8 patients with DNOAP admitted to Honghui Hospital of Xi'an Jiaotong University from March 2017 to June 2018 were selected as DNOAP group,including 3 males and 5 females,aged 20-66(55.7±3.8)years.From April 2017 to July 2018,the articular cartilage of tibial talus joint,subtalar joint and talus-navicular joint of 8 patients with amputation due to traffic accident or serious trauma in Honghui Hospital of Xi'an Jiaotong University were normal control group,including 4 males and 4 females,aged 19-65(57.6±3.7)years.Cartilage samples from DNOAP group and normal control group were taken.Masson staining and safranine O/fixed green staining were used to observe the histopathological characteristics of cartilage,and the ultrastructural changes of chondrocytes were observed by electron microscopy.Chondrocyte culture was carried out in DNOAP group and normal control group.DCFH-DA probe was used to detect the level of reactive oxygen species in chondrocyte.Western blot was used to detect the expression of receptor activator of nuclear factor-κB ligand(RANKL),osteoprotegerin(OPG),IL-1β,IL-6,TNF-α,aggrecan protein in chondrocyte.Flow cytometry(FCM)was used to detect the apoptotic rate of chondrocyte.Results In DNOAP group,chondrocytes decreased,subchondral bone proliferated,structure disordered and osteoclasts formed in a large number in subchondral bone area.Under transmission electron microscopy,mitochondria swelled,membrane structure was incomplete,arrangement disordered,endoplasmic reticulum was severely swollen and nucleus of DNOAP chondrocytes were observed.The chromatin was partially broken and concentrated at the edge of nuclear membrane.In the normal control group,chondrocyte was found in the cartilage lacuna,with homogeneous staining of cartilage matrix and neat arrangement of subchondral

关 键 词：关节病 神经原性 糖尿病 软骨 软骨细胞 病理学 发病机制 

分 类 号：R6[医药卫生—外科学] R587[医药卫生—临床医学]